3D Tissue Culture Devices and Systems

Abstract

Embodiments disclosed herein are directed to bioreactor devices and systems that allow cultured cells to grow and develop with a three-dimensional aspect. In addition, the bioreactor systems described herein can be used for variation and independent control of environmental factors within the individual sub-wells which can be advantageously co-located in a common chamber. For example, the chemical make-up of a nutrient medium that can flow through a chamber as well as the mechanical force environment within the chamber, including the perfusion flow, shear stress, hydrostatic pressure, and the like, can be independently controlled and maintained for each separate culture chamber of the disclosed systems. Further, the bioreactor systems are designed for easy incorporation into automated systems and minimize or eliminate tubing.

1 . A bioreactor device comprising:
 one or more chambers; and   a lid, the lid comprising one or more integrated flow circuits defined therein, the flow circuits comprising flow channels that direct fluids into the one or more chambers.   
     
     
         2 . The bioreactor device of  claim 1 , wherein the one or more chambers are wells of a microwell plate. 
     
     
         3 . The bioreactor device of  claim 1 , further comprising, for each chamber, an insert, the insert defining within the chamber one or more sub-wells. 
     
     
         4 . The bioreactor device of  claim 1 , wherein the lid comprises a first layer and a second layer, wherein the first layer comprises inputs for connecting each flow circuit to a pump, and wherein the flow circuit is defined within the first and second layer. 
     
     
         5 . The bioreactor device of  claim 4 , wherein the second layer further comprises, for each chamber, an extension on the bottom surface of the second layer that seals the second layer to the one or more chambers. 
     
     
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         21 . The bioreactor device of  claim 1 , wherein the one or more chambers further comprise an analytical imaging chamber to facilitate microscopy or spectrometry analysis in the chamber. 
     
     
         22 . The bioreactor device of  claim 1 , wherein the fluidic circuit comprises an analytical imaging window. 
     
     
         23 . The bioreactor device of  claim 3 , wherein the inserts define two or more sub-wells. 
     
     
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         30 . An adapter to convert microwell plates into three-dimensional culture devices comprising a sealing lid to fit within a well of a microwell plate, the sealing lid comprising an input flow channel and an output flow channel, and an insert defining one or more sub-wells within the well of microwell plate. 
     
     
         31 . The adapter of  claim 30 , wherein the insert comprises a non-porous outer region and a porous region around the sub-wells. 
     
     
         32 . The adapter of  claim 30 , wherein the microplate is a 6, 12, 24, 48, 96, or 384 well-sized microplate. 
     
     
         33 . A bioreactor device comprising:
 a bottom soft layer defining one or more inlet and outlet fluid channels and one or more openings; and   a rigid top layer comprising one or more chamber access ports to align with the one or more openings, the one or more access ports comprising one or more channels in the walls of the access ports.   
     
     
         34 . The bioreactor device of  claim 33 , wherein the device is assembled by sealing the rigid top layer to the bottom soft layer such that the one or access ports are inserted through the one or more openings and the one or more channels in the walls of the access port are aligned with the inlet and outlet fluid channels of the soft layer. 
     
     
         35 . The device of  claim 33 , wherein the access port defines a cylindrical cell culture chamber. 
     
     
         36 . The device of  claim 35 , wherein the one or more channels in the walls of the access port enter the chamber at different heights. 
     
     
         37 . A bioreactor device comprising:
 a hard layer defining one or more cell culture chambers, the one or more cell culture chambers comprising an inlet opening and an outlet opening at a base of the cell culture chamber; and   a soft layer for mounting to the hard layer, the soft layer defining one or more openings to receive with the one or more cell culture chambers and inlet and outlet flow channels that align with the inlet and outlet openings at the base of the cell culture chamber.   
     
     
         38 . The device of  claim 37 , wherein each cell culture chamber is connected to the other cell culture chambers by an inlet and outlet flow channel. 
     
     
         39 . A method of culturing one or more cell types in a three-dimensional environment comprising culturing the one or more cell types in a bioreactor device of any one of  claim 1 ,  33 , or  37 . 
     
     
         40 . The method of  claim 39 , wherein one of the one or more cell types is a cancer cell. 
     
     
         41 . The method of  claim 40 , wherein the one or more cell types further comprises epithelial cells, fibroblasts, adipocytes, endothelial cells, tumor associate macrophages, T-cells, B-cells or a combination thereof. 
     
     
         42 . The method of  claim 40 , wherein the cancer cell is a breast cancer cell, a lung cancer cell, ovarian cancer cell, a pancreatic cancer cell, or a glioblastoma. 
     
     
         43 . A method for assessing responsiveness to therapeutic agents comprising:
 culturing in a three-dimensional environment a plurality of cell samples in a plurality of wells or sub-wells of the bioreactor devices of any one of  claim 1 ,  33 , or  37 ;   exposing each cell sample in a well to a different therapeutic agent or a different concentration of the same therapeutic agent by perfusing media through the plurality of chambers comprising the appropriate concentration of therapeutic agent; and   measuring the responsiveness to each cell sample to the therapeutic agent.   
     
     
         44 . The method of  claim 43 , wherein the cell sample is a cancer cell sample. 
     
     
         45 . The method of  claim 44 , wherein the cancer cell sample is a obtained from a biopsy sample of a subject in need of therapeutic treatment. 
     
     
         46 . The method of  claim 45 , wherein the cancer cell sample is a breast cancer sample, ovarian cancer sample, a pancreatic cancer sample, a lung cancer sample, or a glioblastoma. 
     
     
         47 . The method of  claim 43 , wherein the plurality of cell samples is cultured in a microtumor model comprising epithelial cells, fibroblasts, adipocytes, endothelial cells, tumor associate macrophages, T-cells, or B-cells or a combination thereof. 
     
     
         48 . The method of  claim 43 , wherein responsiveness is measured by observable phenotypic changes in cell structure on the device, the release of one or more biomarkers in the perfused media, or through lysis and detection of one or more biomarkers.